U.S. patent number 11,348,772 [Application Number 17/259,580] was granted by the patent office on 2022-05-31 for sample support, sample ionization method, and mass spectrometry method.
This patent grant is currently assigned to HAMAMATSU PHOTONICS K.K.. The grantee listed for this patent is HAMAMATSU PHOTONICS K.K.. Invention is credited to Masahiro Kotani, Takayuki Ohmura, Miu Takimoto.
United States Patent |
11,348,772 |
Takimoto , et al. |
May 31, 2022 |
Sample support, sample ionization method, and mass spectrometry
method
Abstract
A sample support is a sample support for sample ionization,
including: a substrate formed with a plurality of through holes
opening to a first surface and a second surface on a side opposite
to the first surface; a conductive layer provided not to block the
through hole in the first surface; and a frame body provided in a
peripheral portion of the substrate to surround an ionization
region in which a sample is ionized when viewed in a thickness
direction of the substrate, in which a marker for recognizing a
position in the ionization region is provided in the frame
body.
Inventors: |
Takimoto; Miu (Hamamatsu,
JP), Ohmura; Takayuki (Hamamatsu, JP),
Kotani; Masahiro (Hamamatsu, JP) |
Applicant: |
Name |
City |
State |
Country |
Type |
HAMAMATSU PHOTONICS K.K. |
Hamamatsu |
N/A |
JP |
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|
Assignee: |
HAMAMATSU PHOTONICS K.K.
(Hamamatsu, JP)
|
Family
ID: |
1000006342050 |
Appl.
No.: |
17/259,580 |
Filed: |
July 25, 2019 |
PCT
Filed: |
July 25, 2019 |
PCT No.: |
PCT/JP2019/029289 |
371(c)(1),(2),(4) Date: |
January 12, 2021 |
PCT
Pub. No.: |
WO2020/031729 |
PCT
Pub. Date: |
February 13, 2020 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20210319991 A1 |
Oct 14, 2021 |
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Foreign Application Priority Data
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Aug 6, 2018 [JP] |
|
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JP2018-147887 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
H01J
49/0418 (20130101); H01J 49/0004 (20130101) |
Current International
Class: |
H01J
49/00 (20060101); H01J 49/04 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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3686590 |
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Jul 2020 |
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EP |
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2014-021048 |
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Feb 2014 |
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JP |
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6093492 |
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Mar 2017 |
|
JP |
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WO-2008/068847 |
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Jun 2008 |
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WO |
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WO-2017/038710 |
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Mar 2017 |
|
WO |
|
Other References
International Preliminary Report on Patentability dated Feb. 18,
2021 for PCT/JP2019/029289. cited by applicant.
|
Primary Examiner: Ippolito; Nicole M
Assistant Examiner: Chang; Hanway
Attorney, Agent or Firm: Faegre Drinker Biddle & Reath
LLP
Claims
The invention claimed is:
1. A sample support for sample ionization, comprising: a substrate
formed with a plurality of through holes opening to a first surface
and a second surface on a side opposite to the first surface; a
conductive layer provided not to block the through hole in the
first surface; and a frame body provided in a peripheral portion of
the substrate to surround an ionization region in which a sample is
ionized when viewed in a thickness direction of the substrate,
wherein a marker for recognizing a position in the ionization
region is provided in the frame body.
2. The sample support according to claim 1, wherein a width of the
through hole is 1 nm to 700 nm, and a thickness of the substrate is
1 .mu.m to 50 .mu.m.
3. The sample support according to claim 1, wherein a plurality of
first markers disposed along a first direction are provided in a
portion of the frame body extending along the first direction, and
a plurality of second markers disposed along a second direction
orthogonal to the first direction are provided in a portion of the
frame body extending along the second direction.
4. The sample support according to claim 1, wherein the marker is
at least one selected from a numeric character, a signal, and a
letter.
5. The sample support according to claim 1, wherein the marker
includes a marker for visual contact having a width of greater than
or equal to a predetermined value and a marker for a device having
a width of less than the predetermined value.
6. A sample ionization method of an ionization device including an
irradiation unit configured to apply an energy ray, a scanning unit
configured to scan the marker provided in the frame body, and a
control unit configured to control an operation of the irradiation
unit, the method comprising: a first step of preparing a sample and
the sample support according to claim 1; a second step of disposing
the sample support on the sample such that the second surface faces
the sample; a third step of causing the control unit to recognize
an irradiation range of the energy ray in the ionization region by
causing the scanning unit to scan the marker provided in the frame
body; and a fourth step of ionizing a component of the sample moved
to the first surface side through the through hole in the
irradiation range by causing the control unit to operate the
irradiation unit such that the first surface in the irradiation
range is irradiated with the energy ray while a voltage is applied
to the conductive layer.
7. The sample ionization method according to claim 6, wherein the
marker includes a marker for visual contact having a width of
greater than or equal to a predetermined value and a marker for a
device having a width of less than the predetermined value, and in
the third step, a measurer determines the irradiation range, on the
basis of an existence range of the sample in the ionization region
and the marker for visual contact, and the control unit recognizes
the irradiation range, on the basis of a position of the scanning
unit when the marker for a device corresponding to the irradiation
range determined by the measurer is read by the scanning unit.
8. A mass spectrometry method, comprising: each of the steps of the
sample ionization method according to any one of claim 7; a fifth
step of detecting the ionized component and of acquiring a
distribution image indicating a mass distribution of the sample in
the irradiation range; a sixth step of acquiring an optical image
including the sample and the sample support, in a state in which
the sample support is disposed on the sample; and a seventh step of
superimposing the optical image on the distribution image such that
the irradiation range of the optical image overlaps with the
distribution image, on the basis of the marker in the optical
image.
9. A mass spectrometry method, comprising: each of the steps of the
sample ionization method according to claim 6; a fifth step of
detecting the ionized component and of acquiring a distribution
image indicating a mass distribution of the sample in the
irradiation range; a sixth step of acquiring an optical image
including the sample and the sample support, in a state in which
the sample support is disposed on the sample; and a seventh step of
superimposing the optical image on the distribution image such that
the irradiation range of the optical image overlaps with the
distribution image, on the basis of the marker in the optical
image.
10. A sample support for sample ionization, comprising: a substrate
having conductivity, and formed with a plurality of through holes
opening to a first surface and a second surface on a side opposite
to the first surface a frame body provided in a peripheral portion
of the substrate to surround an ionization region in which a sample
is ionized when viewed in a thickness direction of the substrate,
wherein a marker for recognizing a position in the ionization
region is provided in the frame body.
11. A sample ionization method of an ionization device including an
irradiation unit configured to apply an energy ray, a scanning unit
configured to scan the marker provided in the frame body, and a
control unit configured to control an operation of the irradiation
unit, the method comprising: a first step of preparing a sample and
the sample support according to claim 6; a second step of disposing
the sample support on the sample such that the second surface faces
the sample; a third step of causing the control unit to recognize
an irradiation range of the energy ray in the ionization region by
causing the scanning unit to scan the marker provided in the frame
body; and a fourth step of ionizing a component of the sample moved
to the first surface side through the through hole in the
irradiation range by causing the control unit to operate the
irradiation unit such that the first surface in the irradiation
range is irradiated with the energy ray while a voltage is applied
to the substrate.
12. The sample ionization method according to claim 11, wherein the
marker includes a marker for visual contact having a width of
greater than or equal to a predetermined value and a marker for a
device having a width of less than the predetermined value, and in
the third step, a measurer determines the irradiation range, on the
basis of an existence range of the sample in the ionization region
and the marker for visual contact, and the control unit recognizes
the irradiation range, on the basis of a position of the scanning
unit when the marker for a device corresponding to the irradiation
range determined by the measurer is read by the scanning unit.
13. A mass spectrometry method, comprising: each of the steps of
the sample ionization method according to any one of claim 11; a
fifth step of detecting the ionized component and of acquiring a
distribution image indicating a mass distribution of the sample in
the irradiation range; a sixth step of acquiring an optical image
including the sample and the sample support, in a state in which
the sample support is disposed on the sample; and a seventh step of
superimposing the optical image on the distribution image such that
the irradiation range of the optical image overlaps with the
distribution image, on the basis of the marker in the optical
image.
14. A mass spectrometry method, comprising: each of the steps of
the sample ionization method according to any one of claim 12; a
fifth step of detecting the ionized component and of acquiring a
distribution image indicating a mass distribution of the sample in
the irradiation range; a sixth step of acquiring an optical image
including the sample and the sample support, in a state in which
the sample support is disposed on the sample; and a seventh step of
superimposing the optical image on the distribution image such that
the irradiation range of the optical image overlaps with the
distribution image, on the basis of the marker in the optical
image.
Description
TECHNICAL FIELD
The present disclosure relates to a sample support, a sample
ionization method, and a mass spectrometry method.
BACKGROUND ART
In the related art, a sample support for ionizing a sample is known
in mass spectrometry of a sample such as a biological sample (for
example, refer to Patent Literature 1). Such a sample support
includes a substrate formed with a plurality of through holes
opening to a first surface and a second surface on a side opposite
to the first surface. In a case where the sample support is
disposed on the sample such that the second surface faces the
sample, it is possible to lift up the sample from the second
surface side of the substrate toward the first surface side through
the through hole by using a capillary action. Then, in a case where
the first surface side, for example, is irradiated with an energy
ray such as laser beam, the sample moved to the first surface side
is ionized.
CITATION LIST
Patent Literature
Patent Literature 1: Japanese Patent No. 6093492
SUMMARY OF INVENTION
Technical Problem
In the mass spectrometry as described above, when the first surface
side of the substrate is irradiated with the energy ray, it is
required that a mass spectrometry device recognizes an irradiation
range of the energy ray. However, a visual field of a camera or the
like that is attached to the mass spectrometry device is narrow in
accordance with the mass spectrometry device, and it is not
possible to observe the entire sample support disposed in the mass
spectrometry device, and thus, it may not be possible to easily
recognize the irradiation range.
Therefore, an object of one aspect of the present disclosure is to
provide a sample support, a sample ionization method, and a mass
spectrometry method in which an irradiation range of an energy ray
can be easily recognized.
Solution to Problem
A sample support according to one aspect of the present disclosure
is a sample support for sample ionization, including: a substrate
formed with a plurality of through holes opening to a first surface
and a second surface on a side opposite to the first surface; a
conductive layer provided not to block the through hole in the
first surface; and a frame body provided in a peripheral portion of
the substrate to surround an ionization region in which a sample is
ionized when viewed in a thickness direction of the substrate, in
which a marker for recognizing a position in the ionization region
is provided in the frame body.
In the sample support, the plurality of through holes opening to
the first surface and the second surface on a side opposite to the
first surface are formed on the substrate. For this reason, for
example, in a case where the sample support is disposed on a sample
such as a biological sample such that the second surface of the
substrate faces the sample, it is possible to move the sample (a
component of the sample) toward the first surface side from the
second surface side through the through hole by using a capillary
action. Further, for example, in a case where the first surface is
irradiated with an energy ray such as laser beam, energy is
transmitted to the component of the sample moved to the first
surface side via the conductive layer, and thus, it is possible to
ionize the component of the sample. In addition, the sample support
includes the frame body provided in the peripheral portion of the
substrate. For this reason, it is possible to improve the
handleability of the sample support by the frame body. In addition,
the frame body surrounds the ionization region in which the sample
is ionized when viewed in the thickness direction of the substrate,
and the marker for recognizing the position in the ionization
region is provided in the frame body. Accordingly, the following
effects are obtained. That is, for example, a visual field of a
camera or the like that is attached to an ionization device
irradiating the sample support with an energy ray is narrow, and it
may be difficult to specify an irradiation range (a range to be
irradiated with the energy ray) by the observation of the
ionization region. Even in such a case, it is possible for the
ionization device to recognize the irradiation range of the energy
ray, by causing the camera or the like to perform scanning and by
reading the marker provided in the frame body. Accordingly,
according to such a sample support, it is possible to easily
recognize the irradiation range of the energy ray.
A width of the through hole may be 1 nm to 700 nm, and a thickness
of the substrate may be 1 .mu.m to 50 .mu.m. In this case, it is
possible to suitably attain the movement of the component of the
sample by the capillary action described above.
A plurality of first markers disposed along a first direction may
be provided in a portion of the frame body extending along the
first direction, and a plurality of second markers disposed along a
second direction orthogonal to the first direction may be provided
in a portion of the frame body extending along the second
direction. In this case, it is possible to recognize the position
in the first direction by the first marker and to recognize the
position in the second direction by the second marker. Accordingly,
it is possible to easily grasp two-dimensional coordinates of the
irradiation range of the energy ray (for example, a start point
position, an end point position, and the like).
The marker may be at least one selected from a numeric character, a
signal, and a letter. In this case, it is possible to attain the
marker suitable for visual contact and/or for reading a device.
The marker may include a marker for visual contact having a width
of greater than or equal to a predetermined value and a marker for
a device having a width of less than the predetermined value. In
this case, for example, it is possible for a measurer to determine
in advance the irradiation range by visually reading the marker for
visual contact. Further, for example, the marker for a device
corresponding to the irradiation range determined by the measurer
is read by the camera that is attached to the ionization device,
and thus, it is possible for the ionization device to recognize the
irradiation range of the energy ray.
A sample support according to another aspect of the present
disclosure is a sample support for sample ionization, including: a
substrate having conductivity, and formed with a plurality of
through holes opening to a first surface and a second surface on a
side opposite to the first surface; and a frame body provided in a
peripheral portion of the substrate to surround an ionization
region in which a sample is ionized when viewed in a thickness
direction of the substrate, in which a marker for recognizing a
position in the ionization region is provided in the frame
body.
According to such a sample support, it is possible to omit the
conductive layer and to obtain the same effects as those of the
sample support including the conductive layer described above.
A sample ionization method according to one aspect of the present
disclosure is a sample ionization method of an ionization device
including an irradiation unit configured to apply an energy ray, a
scanning unit configured to scan a marker provided in a frame body,
and a control unit configured to control an operation of the
irradiation unit, the method including: a first step of preparing a
sample and the sample support including the conductive layer; a
second step of disposing the sample support on the sample such that
the second surface faces the sample; a third step of causing the
control unit to recognize an irradiation range of the energy ray in
the ionization region by causing the scanning unit to scan the
marker provided in the frame body; and a fourth step of ionizing a
component of the sample moved to the first surface side through the
through hole in the irradiation range by causing the control unit
to operate the irradiation unit such that the first surface in the
irradiation range is irradiated with the energy ray while a voltage
is applied to the conductive layer.
In the sample ionization method described above, the plurality of
through holes opening to the first surface and the second surface
on a side opposite to the first surface are formed on the
substrate. In a case where the sample support is disposed on the
sample such that the second surface of the substrate faces the
sample, the sample (the component of the sample) is moved toward
the first surface side from the second surface side through the
through hole by a capillary action. Further, in a case where the
first surface is irradiated with the energy ray while a voltage is
applied to the conductive layer, energy is transmitted to the
component of the sample moved to the first surface side.
Accordingly, the component of the sample is ionized. In addition,
it is possible for the ionization device to easily recognize the
irradiation range of the energy ray by scanning the marker provided
in the frame body.
A sample ionization method according to another aspect of the
present disclosure is a sample ionization method of an ionization
device including an irradiation unit configured to apply an energy
ray, a scanning unit configured to scan a marker provided in a
frame body, and a control unit configured to control an operation
of the irradiation unit, the method including: a first step of
preparing a sample and the sample support including the substrate
having conductivity; a second step of disposing the sample support
on the sample such that the second surface faces the sample; a
third step of causing the control unit to recognize an irradiation
range of the energy ray in the ionization region by causing the
scanning unit to scan the marker provided in the frame body; and a
fourth step of ionizing a component of the sample moved to the
first surface side through the through hole in the irradiation
range by causing the control unit to operate the irradiation unit
such that the first surface in the irradiation range is irradiated
with the energy ray while a voltage is applied to the
substrate.
According to such a sample ionization method, it is possible to
omit the conductive layer from the sample support and to obtain the
same effects as those in the case of using the sample support
including the conductive layer as described above.
In the ionization method described above, the marker may include a
marker for visual contact having a width of greater than or equal
to a predetermined value and a marker for a device having a width
of less than the predetermined value, and in the third step, a
measurer may determine the irradiation range, on the basis of an
existence range of the sample in the ionization region and the
marker for visual contact, and the control unit may recognize the
irradiation range, on the basis of a position of the scanning unit
when the marker for a device corresponding to the irradiation range
determined by the measurer is read by the scanning unit. In this
case, it is possible to accurately attain both of the determination
of the irradiation range by the visual contact of the measurer and
the recognition of the irradiation range by a mechanical
manipulation (marker scanning) of the ionization device, by the
marker provided in the frame body.
A mass spectrometry method, includes: each of the steps of the
sample ionization method according to one aspect of the present
disclosure described above; a fifth step of detecting the ionized
component and of acquiring a distribution image indicating a mass
distribution of the sample in the irradiation range; a sixth step
of acquiring an optical image including the sample and the sample
support, in a state in which the sample support is disposed on the
sample; and a seventh step of superimposing the optical image on
the distribution image such that the irradiation range of the
optical image overlaps with the distribution image, on the basis of
the marker in the optical image.
According to the mass spectrometry method described above, it is
possible to accurately superimpose the optical image of the sample
on the distribution image, on the basis of the marker provided in
the frame body of the sample support. As a result thereof, it is
possible to visualize the mass distribution in each position of the
sample.
Advantageous Effects of Invention
According to one aspect of the present disclosure, it is possible
to provide a sample support, a sample ionization method, and a mass
spectrometry method in which an irradiation range of an energy ray
can be easily recognized.
BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 is a plan view of a sample support according to one
embodiment.
FIG. 2 is a sectional view of the sample support along line II-II
illustrated in FIG. 1.
FIG. 3 is a diagram illustrating an enlarged image of an effective
region in a substrate viewed in a thickness direction of the
substrate illustrated in FIG. 1.
FIG. 4 is an enlarged view of a frame illustrated in FIG. 1.
FIG. 5 is a diagram illustrating a procedure of a mass spectrometry
method according to one embodiment.
FIG. 6 is a diagram illustrating the procedure of the mass
spectrometry method according to one embodiment.
FIG. 7 is a diagram illustrating the procedure of the mass
spectrometry method according to one embodiment.
DESCRIPTION OF EMBODIMENTS
Hereinafter, preferred embodiments of the present disclosure will
be described in detail, with reference to the drawings. Note that,
in each of the drawings, the same reference numerals will be
applied to the same portions or the corresponding portions, and the
repeated description will be omitted. In addition, dimensions or
dimensional ratios of each member (or part) illustrated in the
drawings may be different from actual dimensions or dimensional
ratios in order to make the description easy to understand.
Configuration of Sample Support
FIG. 1 is a plan view of a sample support 1 of one embodiment. As
illustrated in FIG. 1 and FIG. 2, the sample support 1 includes a
substrate 2, a frame (a frame body) 3, and a conductive layer 4.
The sample support 1 is a sample support for sample ionization. The
sample support 1, for example, is used for ionizing a component of
a sample that is a measurement target, at the time of performing
mass spectrometry.
The substrate 2 includes a first surface 2a and a second surface 2b
on a side opposite to the first surface 2a. A plurality of through
holes 2c are formed on the substrate 2 uniformly (with a
homogeneous distribution). Each of the through holes 2c extends in
a thickness direction of the sample support 1 (that is, the
substrate 2) (hereinafter, simply referred to as a "thickness
direction") and opens to the first surface 2a and the second
surface 2b. The thickness direction is a direction perpendicular to
the first surface 2a and the second surface 2b. The substrate 2,
for example, is formed of an insulating material in a rectangular
plate shape. The length of one side of the substrate 2 when viewed
in the thickness direction, for example, is approximately several
cm to several tens of cm. The thickness of the substrate 2, for
example, is approximately 1 .mu.m to 50 .mu.m. In this embodiment,
the thickness of the substrate 2 is approximately 5 .mu.m. The
substrate 2 is approximately transparent with respect to visible
light. For example, the sample described above can be visually
recognized via the substrate 2.
The frame 3 is provided on the first surface 2a of the substrate 2.
Specifically, the frame 3 is fixed to the first surface 2a of the
substrate 2 by an adhesive layer 5. It is preferable to use an
adhesive material having less emitted gas (for example, low-melting
glass, a vacuum adhesive agent, or the like), as the material of
the adhesive layer 5. The frame 3 has a rectangular frame shape.
The frame 3 is provided in a peripheral portion of the substrate 2.
The frame 3 includes a rectangular inner edge 3a and a rectangular
outer edge 3b. The frame 3 surrounds an effective region (an
ionization region) R when viewed in the thickness direction. The
effective region R is a region that functions in order to move the
component of the sample described below to the first surface 2a
side in the substrate 2 and to ionize the component of the
sample.
The frame 3 has approximately the same outer shape as that of the
substrate 2 when viewed in the thickness direction. The length of
one side of the frame 3 when viewed in the thickness direction (the
length of one side of the outer edge 3b), for example, is
approximately several cm to several tens of cm. The length of one
side of the inner edge 3a (the effective region R) of the frame 3
when viewed in the thickness direction, for example, is
approximately several cm to several tens of cm. The thickness of
the frame 3, for example, is less than or equal to 1 mm. The
material of the frame 3, for example, is a metal, a ceramic, and
the like. According to such a frame 3, the handling of the sample
support 1 is facilitated, and the modification of the substrate 2
due to a temperature change or the like is suppressed.
A first marker 50 and a second marker 60 are provided on a surface
3c of the frame 3 on a side opposite to the substrate 2. A
plurality of first markers 50 are disposed along an X axis
direction in a first portion 31 of the frame 3 extending along the
X axis direction (a first direction). The first marker 50, for
example, is a plurality of numeric characters arranged along the X
axis direction. Similarly, a plurality of second markers 60 are
disposed along a Y axis direction in a second portion 32 of the
frame 3 extending along the Y axis direction (a second direction
orthogonal to the first direction). The second marker 60, for
example, is a plurality of numeric characters arranged along the Y
axis direction. The first marker 50 and the second marker 60
configure a coordinate system for recognizing a position in the
effective region R when viewed in the thickness direction. The
first marker 50 and the second marker 60, for example, are formed
by making the surface 3c of the frame 3 concave and convex.
The conductive layer 4 is provided on the first surface 2a of the
substrate 2. Specifically, the conductive layer 4 is continuously
(integrally) formed in a region corresponding to the inner edge 3a
of the frame 3 (that is, a region corresponding to the effective
region R) on the first surface 2a of the substrate 2, an inner
surface of the inner edge 3a, and the surface 3c of the frame 3. In
the effective region R, the conductive layer 4 is provided in a
peripheral portion of the through hole 2c on the first surface 2a.
That is, the conductive layer 4 covers a portion in the first
surface 2a of the substrate 2, on which the through hole 2c is not
formed. That is, the conductive layer 4 is provided not to block
the through hole 2c. In the effective region R, each of the through
holes 2c is exposed to the inner edge 3a. The conductive layer 4
covers the first marker 50 and the second marker 60 on the surface
3c of the frame 3. However, the first marker 50 and the second
marker 60 are formed by making the surface 3c concave and convex,
and thus, the visual contact and the recognition of the device are
not hindered even in the case of being covered with the conductive
layer 4.
The conductive layer 4 is formed of a conductive material. Here, it
is preferable that a metal having low affinity (reactivity) with
respect to a sample and high conductivity is used as the material
of the conductive layer 4, from the following reasons.
For example, in a case where the conductive layer 4 is formed of a
metal such as copper (Cu) having high affinity with respect to a
sample such as protein, in a process of ionizing the sample
described below, the sample is ionized in a state where Cu atoms
are attached to sample molecules, and thus, there is a concern that
a detection result is shifted in a mass spectrometry method
described below as the Cu atoms are attached. Therefore, it is
preferable that a metal having low affinity with respect to the
sample is used as the material of the conductive layer 4.
On the other hand, a metal having high conductivity easily and
stably applies a constant voltage. For this reason, in a case where
the conductive layer 4 is formed of the metal having high
conductivity, it is possible to homogeneously apply a voltage to
the first surface 2a of the substrate 2. In addition, there is a
tendency that the metal having high conductivity also has high
thermal conductivity. For this reason, in a case where the
conductive layer 4 is formed of the metal having high conductivity,
it is possible to efficiently transfer the energy of an energy ray
such as laser beam that is applied to the substrate 2 to the sample
via the conductive layer 4. Therefore, it is preferable that the
metal having high conductivity is used as the material of the
conductive layer 4.
From the viewpoint described above, for example, it is preferable
that gold (Au), platinum (Pt), and the like are used as the
material of the conductive layer 4. The conductive layer 4, for
example, is formed to have a thickness of approximately 1 nm to 350
nm by a plating method, an atomic layer deposition (ALD) method, an
evaporation method, a sputtering method, and the like. In this
embodiment, the thickness of the conductive layer 4 is
approximately 10 nm. Note that, for example, chromium (Cr), nickel
(Ni), titanium (Ti), and the like may be used as the material of
the conductive layer 4.
FIG. 3 is a diagram illustrating an enlarged image of the substrate
2 when viewed in the thickness direction. In FIG. 3, a black
portion is the through hole 2c, and a white portion is a partition
portion between the through holes 2c. As illustrated in FIG. 3, the
plurality of through holes 2c having an approximately constant
width are uniformly formed on the substrate 2. The through hole 2c,
for example, is approximately in a circle shape when viewed in the
thickness direction. The width of the through hole 2c, for example,
is approximately 1 nm to 700 nm. In this embodiment, the width of
the through hole 2c is approximately 200 nm. The width of the
through hole 2c indicates the diameter of the through hole 2c in a
case where the through hole 2c is approximately in a circle shape
when viewed in the thickness direction, and indicates the diameter
(an effective diameter) of a virtual maximum cylinder falling into
the through hole 2c in a case where the through hole 2c is not
approximately in a circle shape. A pitch between the respective
through holes 2c, for example, is approximately 1 nm to 1000 nm. In
a case where the through hole 2c is approximately in a circle shape
when viewed in the thickness direction, the pitch between the
respective through holes 2c indicates a center-to-center distance
of the respective circles, and in a case where the through hole 2c
is not approximately in a circle shape, the pitch between the
respective through holes 2c indicates a center axis-to-center axis
distance of the virtual maximum cylinder falling into the through
hole 2c. The width of the partition portion between the through
holes 2c on the substrate 2, for example is approximately 300
nm.
An opening rate of the through holes 2c (a ratio of all of the
through holes 2c to the first surface 2a when viewed in the
thickness direction) is practically 10% to 80%, and is particularly
preferably 50% to 80%. The sizes of the plurality of through holes
2c may be uneven with each other, and the plurality of through
holes 2c may be partially connected to each other.
The substrate 2, for example, is an alumina porous film that is
formed by performing anodic oxidation with respect to aluminum
(Al). Specifically, an anodic oxidation treatment is performed with
respect to an Al substrate, and a surface portion that is oxidized
is peeled off from the Al substrate, and thus, the substrate 2 can
be obtained. Note that, the substrate 2 may be formed by performing
anodic oxidation with respect to a valve metal other than Al, such
as tantalum (Ta), niobium (Nb), titanium (Ti), hafnium (Hf),
zirconium (Zr), zinc (Zn), tungsten (W), bismuth (Bi), and antimony
(Sb), or may be formed by performing anodic oxidation with respect
to silicon (Si).
FIG. 4 is an enlarged view of the frame 3. As illustrated in FIG.
4, the first marker 50 includes a marker 51 for visual contact and
a marker 52 for a device. The marker 51 for visual contact is a
marker to be read by the visual contact of a measurer. A plurality
of markers 51 for visual contact are disposed along the X axis
direction. The plurality of markers 51 for visual contact, for
example, are arranged at regular intervals along the X axis
direction. In this embodiment, as an example, each of the markers
51 for visual contact is a numeric character. A width w1 of the
marker 51 for visual contact can be recognized by the visual
contact of the measurer. The width w1 of the marker 51 for visual
contact is greater than or equal to a predetermined value (for
example, 1 mm).
The width w1 of the marker 51 for visual contact, for example, is
approximately 1 mm to 4 mm. A pitch w2 between the markers 51 for
visual contact adjacent to each other (that is, a distance between
the centers of the markers 51 for visual contact adjacent to each
other), for example, is approximately 2 mm to 10 mm. A length w3 of
a space between the markers 51 for visual contact adjacent to each
other, for example, is approximately 1 mm to 9 mm. In this
embodiment, as an example, the width w1 of the marker 51 for visual
contact is approximately 1 mm, the pitch w2 between the markers 51
for visual contact is approximately 2 mm, and the length w3 of the
space between the markers 51 for visual contact is approximately 1
mm. The marker 51 for visual contact, for example, is provided by
forming concavities and convexities on the surface 3c of the frame
3, in accordance with a punch mark using stamping or laser. An
engraved height (a depth) of the marker 51 for visual contact, for
example, is approximately 0.1 mm to 0.9 mm.
The marker 52 for a device, for example, is a marker to be read by
a camera 16 (refer to FIG. 7) that is attached to a mass
spectrometry device 10 (an ionization device) described below. In
this embodiment, as an example, the marker 52 for a device is
positioned inside the frame 3 from the marker 51 for visual
contact. A plurality of markers 52 for a device are disposed along
the X axis direction. The plurality of markers 52 for a device, for
example, are arranged at regular intervals along the X axis
direction. In this embodiment, as an example, each of the markers
52 for a device is a numeric character.
A width w4 of the marker 52 for a device is less than the width w1
of the marker 51 for visual contact. That is, the width w4 of the
marker 52 for a device is less than a predetermined value. The
width w4 of the marker 52 for a device, for example, is
approximately 1 nm to 0.1 mm. A pitch w5 between the markers 52 for
a device adjacent to each other (that is, a distance between the
centers of the markers 52 for a device adjacent to each other) is
less than the pitch w2 of the marker 51 for visual contact. The
pitch w5 between the markers 52 for a device, for example, is
approximately 5 nm to 0.2 mm. A length w6 of a space between the
markers 52 for a device adjacent to each other, for example, is
approximately 4 nm to 0.1 mm. In this embodiment, as an example,
the width w4 of the marker 52 for a device is approximately 40
.mu.m, the pitch w5 between the markers 52 for a device is
approximately 110 .mu.m, and the length w6 of the space between the
markers 52 for a device is approximately 70 .mu.m. The marker 52
for a device, for example, is provided by forming concavities and
convexities on the surface 3c of the frame 3, in accordance with a
punch mark (as an example, shallow engraving) using laser. An
engraved height (a depth) of the marker 52 for a device, for
example, is approximately 5 .mu.m. Note that, as the punch mark
using the laser, for example, a method such as deep engraving may
be applied. The engraved height of the marker 52 for a device is
different in accordance with each method.
Note that, a positional relationship between the plurality of
markers 51 for visual contact and the plurality of markers 52 for a
device, for example, is stored in a correspondence table that is
prepared in advance. The measurer is capable of grasping that a
marker "14" or "15" of the marker 52 for a device corresponds to a
center position of a marker "1" of the marker 51 for visual contact
in the X axis direction, or an intermediate position of markers "1"
and "2" of the marker 51 for visual contact in the X axis direction
corresponds to a marker "22" of the marker 52 for a device, with
reference to such a correspondence table. Note that, the
correspondence table may be saved on paper or the like, or may be
saved in a storage device (a memory, a storage, or the like) of a
computer as data.
As with the first marker 50, the second marker 60 also includes a
marker 61 for visual contact (refer FIG. 1) and a marker 62 for a
device (refer FIG. 1). That is, the marker 61 for visual contact is
a marker to be read by the visual contact of the measurer. A
plurality of markers 61 for visual contact are disposed along the Y
axis direction. The plurality of markers 61 for visual contact, for
example, are arranged at regular intervals along the Y axis
direction. In this embodiment, as an example, each of the markers
61 for visual contact is a numeric character. The width of the
marker 61 for visual contact, a pitch between the adjacent markers
61 for visual contact, and the length of a space between the
adjacent markers 61 for visual contact are identical to the width
w1 of the marker 51 for visual contact, the pitch w2 between the
adjacent markers 51 for visual contact, and the length w3 of the
space between the adjacent markers 51 for visual contact, described
above. The marker 62 for a device, for example, is a marker to be
read by the camera 16 that is attached to the mass spectrometry
device 10. In this embodiment, as an example, the marker 62 for a
device is positioned inside the frame 3 from the marker 61 for
visual contact. The plurality of markers 62 for a device are
disposed along the Y axis direction. The plurality of markers 62
for a device, for example, are arranged at regular intervals along
the Y axis direction. In this embodiment, as an example, each of
the markers 62 for a device is a numeric character. The width of
the marker 62 for a device, a pitch between the adjacent markers 62
for a device, and the length of a space between the adjacent
markers 62 for a device are identical to the width w4 of the marker
52 for a device, the pitch w5 between the adjacent markers 52 for a
device, and the length w6 of the space between the adjacent markers
52 for a device, described above.
Sample Ionization Method
Next, a sample ionization method using the sample support 1 will be
described with reference to FIG. 5 to FIG. 7. Here, as an example,
a laser desorption/ionization method using laser beam (an energy
ray) (a part of a mass spectrometry method of a mass spectrometry
device 10) will be described. In FIG. 5 and FIG. 7, the through
hole 2c, the conductive layer 4, and the adhesive layer 5 in the
sample support 1 are not illustrated.
First, as illustrated in (a) of FIG. 5, a sample S is prepared (a
first step). Specifically, the sample S is mounted on a mounting
surface 6a of a glass slide (a mounting portion) 6. The glass slide
6 is a glass substrate on which a transparent conductive film such
as an indium tin oxide (ITO) film is formed, and the surface of the
transparent conductive film is the mounting surface 6a. Note that,
not only the glass slide 6 but also a member that is capable of
ensuring conductivity (for example, a substrate formed of a metal
material such as stainless steel, or the like) can be used as the
mounting portion. Here, the sample S, for example, is a biological
sample (a hydrous sample). The sample S, for example, is a liver
slice of a mouse, or the like. In order to smoothly move a
component 51 of the sample S (refer to (c) of FIG. 5), a solution
for decreasing the viscosity of the component 51 (for example, an
acetonitrile mixed liquid, acetone, or the like) may be added to
the sample S.
Subsequently, as illustrated in (b) of FIG. 5, the sample support 1
described above is prepared (the first step). The sample support 1
may be prepared by being manufactured by a person who carries out
the ionization method and the mass spectrometry method, or may be
prepared by being acquired from a manufacturer, a seller, or the
like of the sample support 1. Subsequently, the sample support 1 is
disposed on the sample S such that the second surface 2b faces the
sample S (a second step). The sample support 1 is disposed on the
sample S such that the second surface 2b is in contact with the
sample S.
Subsequently, as illustrated in (c) of FIG. 5, the sample support 1
is fixed to the glass slide 6. The sample support 1 is fixed to the
glass slide 6 by a tape 7 having conductivity (for example, a
carbon tape or the like). The tape 7 fixes the sample support 1
such that the first marker 50 and the second marker 60 are exposed.
That is, the first marker 50 and the second marker 60 are not
covered with the tape 7. Alternatively, for example, the tape 7 is
formed of a transparent material, and thus, in a case where the
first marker 50 and the second marker 60 can be read by the camera
16 described below even when the first marker 50 and the second
marker 60 are covered with the tape 7, the first marker 50 and the
second marker 60 may be covered with the tape 7. The tape 7 may be
a part of the sample support 1, or may be prepared separately from
the sample support 1. In a case where the tape 7 is a part of the
sample support 1 (that is, in a case where the sample support 1
includes the tape 7), for example, the tape 7 may be fixed in
advance to the surface 3c side of the frame 3. More specifically,
the tape 7 may be fixed onto the conductive layer 4 that is formed
on the surface 3c of the frame 3.
The component S1 of the sample S is moved toward the first surface
2a side of the substrate 2 from the second surface 2b side of the
substrate 2 through the through hole 2c by a capillary action. The
component S1 that is moved to the first surface 2a side of the
substrate 2 is accumulated on the first surface 2a side by a
surface tension. FIG. 6 is a plan view illustrating a state in
which the sample support 1 is disposed on the sample S. As
illustrated in FIG. 6, the component S1 of the sample S is moved to
the first surface 2a side in a region D1 of the effective region R
(an existence range of the sample S). Subsequently, the camera 16
scans the first marker 50 and the second marker 60 provided in the
frame 3, and thus, an irradiation range D2 of laser beam L in the
effective region R is recognized by a control unit 17 (refer to
FIG. 7) (a third step).
Specifically, first, the measurer determines the irradiation range
D2, on the basis of the region D1 and the marker 51 for visual
contact and the marker 61 for visual contact. More specifically, in
a case where the sample support 1 is disposed on the sample S, the
measurer, for example, determines a region including the region D1
as the irradiation range D2, by the visual contact. In this
embodiment, as an example, the irradiation range D2 has a rectangle
shape surrounded by a pair of side portions extending in the X axis
direction and a pair of side portions extending in the Y axis
direction. Then, the measurer grasps coordinates (X1,Y1) and
(X2,Y2) of a start point P1 and an end point P2 of the irradiation
range D2, on the basis of the marker 51 for visual contact and the
marker 61 for visual contact, respectively. The coordinates (X1,Y1)
and (X2,Y2) are coordinates in a coordinate system configured of
the marker 51 for visual contact and the marker 61 for visual
contact. Here, as an example, (X1,Y1) is (7.9,1.1), and (X2,Y2) is
(1.5,5.9). Note that, the coordinates that are grasped as described
above are values presumed by the visual contact of the
measurer.
Subsequently, as illustrated in FIG. 7, in a state where the sample
S is disposed between the glass slide 6 and the sample support 1,
the glass slide 6, the sample support 1, and the sample S are
mounted on a support portion 12 of the mass spectrometry device
10.
The mass spectrometry device 10 includes the support portion 12, a
sample stage 18, the camera 16 (a scanning unit), an irradiation
unit 13, a voltage application unit 14, an ion detection unit 15,
and the control unit 17. The sample S or the like that is a
spectrometry target is mounted on the support portion 12. The
support portion 12 on which the sample S or the like is mounted is
mounted on the sample stage 18. The sample S or the like mounted on
the support portion 12 is observed by the camera 16. Here, the
width of an observation range (a visual field) C (refer to FIG. 4)
of the camera 16, for example, is approximately 1.5 mm. That is,
the visual field of the camera 16 is smaller than the effective
region R, and has at least a size capable of observing the markers
52 and 62 for a device. The irradiation unit 13 irradiates the
first surface 2a of the sample support 1 with the energy ray such
as the laser beam L. The voltage application unit 14 applies a
voltage to the first surface 2a of the sample support 1. The ion
detection unit 15 detects ions of the sample S that is ionized. The
control unit 17 controls the operations of the sample stage 18, the
camera 16, the irradiation unit 13, the voltage application unit
14, and the ion detection unit 15. The control unit 17, for
example, is a computer device including a processor (for example, a
central processing unit [CPU]), a memory (for example, a read only
memory [ROM], a random access memory [RAM], or the like), and the
like.
Subsequently, the control unit 17 recognizes irradiation range D2,
on the basis of the marker 52 for a device and the marker 62 for a
device. For example, first, the control unit 17 conveys the sample
support 1 to the sample stage 18 up to a position in which the
marker 52 for a device provided in the frame 3 is imaged by the
camera 16. Specifically, the control unit 17 positions the sample
support 1 such that a target marker 52 for a device falls within
the observation range C of the camera 16 (refer to FIG. 4), by
moving the sample stage 18 such that the camera 16 scans the
markers 52 for a device arrayed along the X axis direction. Here,
the control unit 17 acquires in advance the marker 52 for a device
corresponding to the coordinate X1 (here, 7.9) of the start point
P1 and the coordinate X2 (here, 1.5) of the end point P2, which are
grasped by the visual contact of the measurer. Such acquisition of
the marker 52 for a device, for example, can be attained by an
input manipulation of the measurer with respect to a computer
configuring the control unit 17. Then, the control unit 17
recognizes (stores) the X coordinate of the start point P1 of the
irradiation range D2, on the basis of a position x1 of the sample
stage 18 in the X axis direction when the marker 52 for a device
corresponding to the coordinate X1 of the start point P1 is read by
the camera 16. That is, the control unit 17 recognizes the position
x1 as the X coordinate of the start point P1. Similarly, the
control unit 17 recognizes (stores) the X coordinate of the end
point P2 of the irradiation range D2, on the basis of a position x2
of the sample stage 18 in the X axis direction when the marker 52
for a device corresponding to the coordinate X2 of the end point P2
is read by the camera 16. That is, the control unit 17 recognizes
the position x2 as the X coordinate of the end point P2.
Similarly, the control unit 17 conveys the sample support 1 to the
sample stage 18 up to a position in which the marker 62 for a
device provided in the frame 3 is imaged by the camera 16.
Specifically, the control unit 17 positions the sample support 1
such that a target marker 62 for a device falls within the
observation range C of the camera 16, by moving the sample stage 18
such that the camera 16 scans the markers 62 for a device arrayed
along the Y axis direction. Here, the control unit 17 acquires in
advance the marker 62 for a device corresponding to the coordinate
Y1 (here, 1.1) of the start point P1 and the coordinate Y2 (here,
5.9) of the end point P2, which are grasped by the visual contact
of the measurer. Such acquisition of the marker 62 for a device can
be attained by the same method as the acquisition of the marker 52
for a device described above. Then, the control unit 17 recognizes
(stores) the Y coordinate of the start point P1 of the irradiation
range D2, on the basis of a position y1 of the sample stage 18 in
the Y axis direction when the marker 62 for a device corresponding
to the coordinate Y1 of the start point P1 is read by the camera
16. That is, the control unit 17 recognizes the position y1 as the
Y coordinate of the start point P1. Similarly, the control unit 17
recognizes (stores) the Y coordinate of the end point P2 of the
irradiation range D2, on the basis of a position y2 of the sample
stage 18 in the Y axis direction when the marker 62 for a device
corresponding to the coordinate Y2 of the end point P2 is read by
the camera 16. That is, the control unit 17 recognizes the position
y2 as the Y coordinate of the end point P2.
As described above, the control unit 17 is capable of recognizing
the positions (x1,y1) and (x2,y2) of the start point P1 and the end
point P2, respectively, by causing the camera 16 to scan the
plurality of markers 52 for a device arrayed along the X axis
direction and the plurality of markers 62 for a device arrayed
along the Y axis direction. Accordingly, the control unit 17
recognizes the irradiation range D2. Note that, the coordinates
(X1,Y1) and (X2,Y2) are the coordinates in the coordinate system
configured of the marker 51 for visual contact and the marker 61
for visual contact of the frame 3, whereas the positions (x1,y1)
and (x2,y2) are a control coordinate system that is used by the
control unit 17. That is, the positions (x1,y1) and (x2,y2) are a
position referred to when the irradiation unit 13 performs scanning
Here, the coordinates (X1,Y1) of the start point P1 and the
coordinates (X2,Y2) of the end point P2 correspond to the positions
(x1,y1) and (x2,y2). From such a correspondence relationship, the
coordinate system configured of the marker 51 for visual contact
and the marker 61 for visual contact of the frame 3 and the control
coordinate system can be exchanged each other.
Subsequently, a voltage is applied to the conductive layer 4 of the
sample support 1 (refer to FIG. 2) via the mounting surface 6a of
the glass slide 6 and the tape 7 by the voltage application unit 14
(a fourth step). Subsequently, the control unit 17 operates the
irradiation unit 13, on the basis of irradiation range D2 that is
recognized by the control coordinate system (that is, a range that
is specified by the positions (x1,y1) and (x2,y2)). Specifically,
the control unit 17 operates the irradiation unit 13 such that the
first surface 2a in the irradiation range D2 is irradiated with the
laser beam L (the fourth step). Accordingly, the irradiation unit
13 scans the first surface 2a in the irradiation range D2 with the
laser beam L.
As an example, the control unit 17 moves the sample stage 18, and
controls an irradiation operation (an irradiation timing or the
like) of the laser beam L by the irradiation unit 13, as the
control of the operation of the irradiation unit 13. That is, the
control unit 17 checks that the sample stage 18 is moved by a
predetermined interval, and then, executes the irradiation of the
laser beam L with respect to the irradiation unit 13. Specifically,
first, the control unit 17 moves the sample stage 18, and thus, the
irradiation position of the laser beam L by the irradiation unit 13
is adjusted to the position (x1,y1) corresponding to the start
point P1 of the irradiation range D2. Then, the laser beam L is
applied to the position (x1,y1) of the start point P1.
Subsequently, the control unit 17 moves the sample stage 18, and
thus, the irradiation position of the laser beam L by the
irradiation unit 13 is adjusted to a position separated from the
position (x1,y1) in the X axis direction by a predetermined
interval (a laser irradiation interval set in advance), and the
laser beam L is applied to the position. By repeating such a
manipulation, the laser beam L is sequentially applied at each
predetermined interval in the X axis direction. Then, in a case
where the irradiation position of the laser beam L reaches an edge
portion of the irradiation range D2 (that is, a position (x2,y1)
corresponding to coordinates (X3,Y3) of a turn-around point P3 in
FIG. 6), the control unit 17 moves the sample stage 18, and thus,
the irradiation position of the laser beam L by the irradiation
unit 13 is adjusted to a position separated from the immediately
preceding irradiation position in the Y axis direction by a
predetermined interval, and the laser beam L is applied to the
position. Subsequently, the laser beam L is applied to a position
separated from the immediately preceding irradiation position in
the X axis direction by a predetermined interval. The laser beam L
sequentially applied at each predetermined interval in the X axis
direction. As described above, the laser beam L is scanned in a
meander shape in the irradiation range D2, and then, reaches the
position (x2,y2) of the end point P2. As described above, the first
surface 2a in the irradiation range D2 is scanned with the laser
beam L. Note that, the scanning of the laser beam L with respect to
the first surface 2a can be carried out by operating at least one
of the sample stage 18 and the irradiation unit 13. In a case where
the irradiation unit 13 is operated, the control unit 17 controls
the operation of the irradiation unit 13, and controls both of the
movement of the irradiation unit 13 and the irradiation operation
of the laser beam L by the irradiation unit 13.
As described above, the first surface 2a in the irradiation range
D2 is irradiated with the laser beam L while a voltage is applied
to the conductive layer 4, and thus, the component S1 that is moved
to the first surface 2a side through the through hole 2c in the
irradiation range D2 is ionized, and a sample ion S2 (the component
S1 that is ionized) is emitted. Specifically, energy is transmitted
from the conductive layer 4 absorbing the energy of the laser beam
L to the component S1 that is moved to the first surface 2a side of
the substrate 2, and the component S1 obtaining the energy is
gasified and obtains a charge, and thus, the sample ion S2 is
obtained. Each of the steps described above corresponds to the
ionization method of the sample S, using the sample support 1
(here, as an example, a laser desorption/ionization method as a
part of the mass spectrometry method).
The sample ion S2 that is emitted is moved toward a ground
electrode (not illustrated) that is provided between the sample
support 1 and an ion detection unit 15 while being accelerated.
That is, the sample ion S2 is moved toward the ground electrode
while being accelerated by a potential difference that occurs
between the conductive layer 4 to which a voltage is applied and
the ground electrode. Then, the sample ion S2 is detected by the
ion detection unit 15 (a fifth step).
A detection result of the sample ion S2 by the ion detection unit
15 is associated with the irradiation position of the laser beam L.
Specifically, the ion detection unit 15 detects the sample ion S2
in each position in the irradiation range D2 to which the laser
beam L is applied, as described above. An identification number
indicating each position (that is, the position to which the laser
beam L is applied) (for example, the coordinates or the like in the
control coordinate system, such as (x1,y1) described above) is
applied to the data (the detection result) of the sample ion S2
detected in each position in the irradiation range D2. Accordingly,
a distribution image (MS mapping data) indicating a mass
distribution of the sample S in the irradiation range D2 is
acquired. Further, it is possible to image a two-dimensional
distribution of molecules configuring the sample S. Note that,
here, the mass spectrometry device 10 is a mass spectrometry device
using a time-of-flight mass spectrometry (TOF-MS) method.
Subsequently, in a state where the sample support 1 is disposed on
the sample S, an optical image of the sample S and the sample
support 1 is acquired (a sixth step). Here, the acquired optical
image includes at least of the region D1 in the effective region R,
and the first marker 50 and the second marker 60. Subsequently, the
optical image is superimposed on the distribution image such that
the irradiation range D2 in the optical image overlaps with the
distribution image of the sample S, on the basis of the marker 51
for visual contact and the marker 61 for visual contact in the
optical image (a seventh step). Specifically, the position (x1,y1)
of the start point P1 and the position (x2,y2) of the end point P2
in the distribution image are superimposed on the coordinates
(X1,Y1) of the start point P1 and the coordinates (X2,Y2) of the
end point P2 in the optical image, respectively. Accordingly, the
optical image of the sample S and the distribution image are
synthesized. Note that, the substrate 2 is approximately
transparent with respect to visible light, and thus, even in a case
where the sample support 1 is disposed on the sample S, it is
possible to acquire the optical image of the sample S. Each of the
steps described above corresponds to the mass spectrometry method
using the sample support 1.
After the observation (optical observation) of the optical image of
the sample S and the distribution image of the sample S that are
synthesized together, detailed screening of a liquid
chromatography-mass spectrometry (LC-MS) method may be further
performed with respect to a portion having specific data in the
sample S. In such a case, for example, an identification number
such as the information of the irradiation position is applied to
the specific data, and thus, it is possible to specify the
irradiation position and the coordinates in the sample support 1,
on the basis of the identification number. Accordingly, it is
possible to supply a sample of the sample S in the coordinates to
LC-MS.
As described above, in the sample support 1, the plurality of
through holes 2c opening to the first surface 2a and the second
surface 2b on a side opposite to the first surface 2a are formed on
the substrate 2. For this reason, in a case where the sample
support 1 is disposed on the sample S such that the second surface
2b of the substrate 2 faces the sample S, it is possible to move
the component S1 of the sample S toward the first surface 2a side
from the second surface 2b side through the through hole 2c by
using a capillary action. Further, in a case where the first
surface 2a is irradiated with the laser beam, the energy is
transmitted to the component S1 of the sample S that is moved to
the first surface 2a side via the conductive layer 4, and thus, it
is possible to ionize the component S1 of the sample S. In
addition, the sample support 1 includes the frame 3 provided in the
peripheral portion of the substrate 2. For this reason, it is
possible to improve the handleability of the sample support 1 by
the frame 3. In addition, the frame 3 surrounds the effective
region R in which the sample S is ionized when viewed in the
thickness direction of the substrate 2, and the first marker 50 and
the second marker 60 for recognizing the position in the effective
region R are provided in the frame 3. Accordingly, the following
effects are obtained.
That is, in this embodiment, in a case where the visual field of
the camera 16 is narrow, and the irradiation range D2 is specified
by observing the effective region R (the entire sample S), it is
difficult to determine the irradiation position of the laser beam
L. According to the sample support 1, it is possible for the mass
spectrometry device 10 to recognize the irradiation range D2 of the
laser beam L, by causing the camera 16 to perform scanning and by
reading the first marker 50 and the second marker 60 provided in
the frame 3. Accordingly, according to the sample support 1, it is
possible to easily recognize the irradiation range D2 of the laser
beam L. In particular, in a mass spectrometry device (an MALDI-MS
device) that is used in existing MALDI, the number of pixels, the
visual field, or the like of the camera that is attached to the
device is optimized to be suitable for the observation of matrix
crystals in the sample, and thus, there is a problem that the mass
spectrometry device is not suitable for the determination of the
irradiation position of the laser beam L and the observation of the
entire sample for imaging. It is possible to solve the problem of
the MALDI-MS device described above by applying the mass
spectrometry device 10 described above to such an MALDI-MS
device.
The width of the through hole 2c is 1 nm to 700 nm, and the
thickness of the substrate 2 is 1 .mu.m to 50 .mu.m. Accordingly,
it is possible to suitably attain the movement of the component S1
of the sample S by the capillary action described above.
The plurality of first markers 50 disposed along the X axis
direction are provided in the first portion 31 extending along the
X axis direction of the frame 3, and the plurality of second
markers 60 disposed along the Y axis direction are provided in the
second portion 32 extending along the Y axis direction orthogonal
to the X axis direction of the frame 3. Accordingly, it is possible
to recognize the position in the X axis direction by the first
marker 50 and to recognize the position in the Y axis direction by
the second marker 60. Accordingly, it is possible to easily grasp
two-dimensional coordinates of the irradiation range D2 of the
laser beam L (for example, the position of the start point P1, the
position of the end point P2, or the like).
The first marker 50 and the second marker 60 are a numeric
character. Accordingly, it is possible to attain a marker suitable
for visual contact and/or for reading a device.
The first marker 50 includes the marker 51 for visual contact
having the width w1 of greater than or equal to the predetermined
value and the marker 52 for a device having the width w4 of less
than the predetermined value, and the second marker 60 includes the
marker 61 for visual contact having a width of greater than or
equal to a predetermined value and a marker 62 for a device having
a width of less than the predetermined value. Accordingly, the
markers 51 and 61 for visual contact are read by the visual contact
of the measurer, and thus, it is possible to determine in advance
the irradiation range D2. Further, the markers 52 and 62 for a
device corresponding to the irradiation range D2 determined by the
measurer are read by the camera 16, and thus, it is possible for
the mass spectrometry device 10 to recognize the irradiation range
D2 of the laser beam L.
In addition, in the ionization method of the sample S described
above, the plurality of through holes 2c opening to the first
surface 2a and the second surface 2b on a side opposite to the
first surface 2a are formed on the substrate 2. In a case where the
sample support 1 is disposed on the sample S such that the second
surface 2b of the substrate 2 faces the sample S, the component S1
of the sample S is moved toward the first surface 2a side from the
second surface 2b side through the through hole 2c by the capillary
action. Further, in a case where the first surface 2a is irradiated
with the laser beam L while a voltage is applied to the conductive
layer 4, the energy is transmitted to the component S1 of the
sample S that is moved to the first surface 2a side. Accordingly,
the component S1 of the sample S is ionized. In addition, it is
possible for the mass spectrometry device 10 to easily recognize
the irradiation range D2 of the laser beam L by scanning the first
marker 50 and the second marker 60 provided in the frame 3.
In the ionization method, the first marker 50 includes the marker
51 for visual contact having the width w1 of greater than or equal
to the predetermined value and the marker 52 for a device having
the width w4 of less than the predetermined value, and the second
marker 60 includes the marker 61 for visual contact having the
width of greater than or equal to the predetermined value and the
marker 62 for a device having the width of less than the
predetermined value. Then, in the third step, the measurer
determines the irradiation range D2, on the basis of the region D1
in the effective region R, and the markers 51 and 61 for visual
contact, and the control unit 17 recognizes the irradiation range
D2, on the basis of the position of the sample stage 18 when the
markers 52 and 62 for a device corresponding to the irradiation
range D2 determined by the measurer are read by the camera 16.
Accordingly, it is possible to accurately attain both of the
determination of the irradiation range D2 by the visual contact of
the measurer and the recognition of the irradiation range D2 by a
mechanical manipulation (marker scanning) of the mass spectrometry
device 10, by the first marker 50 and the second marker 60 provided
in the frame 3.
As described above, according to the mass spectrometry method
described above, it is possible to accurately superimpose the
optical image of the sample S on the distribution image, on the
basis of the first marker 50 and the second marker 60 provided in
the frame 3 of the sample support 1. As a result thereof, it is
possible to visualize the mass distribution in each position of the
sample S.
Modification Example
As described above, the embodiment of the present disclosure has
been described, but the present disclosure is not limited to the
embodiment described above, and the present disclosure can be
variously modified within a range not departing from the gist
thereof.
The substrate 2 may have conductivity, and in the mass spectrometry
method, the first surface 2a may be irradiated with the laser beam
L while a voltage is applied to the substrate 2. In a case where
the substrate 2 has conductivity, it is possible to omit the
conductive layer 4 in the sample support 1 and to obtain the same
effects as those in the case of using the sample support 1
including the conductive layer 4 described above. Note that,
irradiating the first surface 2a of the substrate 2 with the laser
beam L indicates that the conductive layer 4 is irradiated with the
laser beam L in a case where the sample support 1 includes the
conductive layer 4, and indicates that the first surface 2a of the
substrate 2 is irradiated with the laser beam L in a case where the
substrate 2 has conductivity.
An example has been described in which the first marker 50 and the
second marker 60 are a numeric character, but the first marker 50
and the second marker 60 may be various markers. The first marker
50 and the second marker 60, for example, may be at least one
selected from a numeric character, a signal, and a letter. Even in
this case, it is possible to attain a marker suitable for visual
contact and/or for reading a device. Each of the marker 51 for
visual contact, the marker 52 for a device, the marker 61 for
visual contact, and the marker 62 for a device, for example, may be
at least one selected from a numeric character, a signal, and a
letter. In addition, such markers may include auxiliary information
such as a graduation line.
An example has been described in which the first marker 50 and the
second marker 60 are formed by making the surface 3c of the frame 3
concave and convex, but the first marker 50 and the second marker
60 may not be formed by making the surface 3c of the frame 3
concave and convex. The first marker 50 and the second marker 60,
for example, may be formed by a print according to printing such as
nanoprinting, photolithography using extreme ultraviolet (EUV)
lithographic exposure, write according to a paint, black
(oxidization) that is an example of a punch mark using a laser,
foamed marking using a laser, or the like. In a case where the
first marker 50 and the second marker 60 are formed by the foamed
marking using the laser, the material of the frame 3 is a resin.
The foamed marking using the laser is a method for foaming the
resin by a laser beam. According to such a method, light is
diffusely reflected in a foamed portion, and as a result thereof,
the visibility of the foamed portion increases. Note that, in a
case where the first marker 50 and the second marker 60 are not
formed by making the surface 3c of the frame 3 concave and convex
and the visibility is inhibited at the time of being covered with
the conductive layer 4, the conductive layer 4 may not be formed in
a region on the surface 3c of the frame 3, in which the first
marker 50 and the second marker 60 are formed.
The frame 3 may include only one of the first marker 50 and the
second marker 60. In this case, it is possible for the mass
spectrometry device 10 to recognize a range in at least one
direction of the irradiation range D2 in the X axis direction and
the Y axis direction. In addition, the first marker 50 may be
provided in both of the first portions 31 facing each other in the
frame 3. Similarly, the second marker 60 may be provided in both of
the second portions 32 facing each other in the frame 3.
The first marker 50 may include only one of the marker 51 for
visual contact and the marker 52 for a device. In this case, it is
preferable that the width of the first marker 50 can be read by any
of the visual contact of the measurer and the camera 16 that is
attached to the mass spectrometry device 10. Similarly, the second
marker 60 may include only one of the marker 61 for visual contact
and the marker 62 for a device. In this case, it is preferable that
the width of the second marker 60 can be read by any of the visual
contact of the measurer and the camera 16 that is attached to the
mass spectrometry device 10.
An example has been described in which the determination of the
irradiation range D2 by the measurer is performed before the glass
slide 6, the sample support 1, and the sample S are mounted on the
support portion 12 of the mass spectrometry device 10, but such
determination of the irradiation range D2 by the measurer may be
performed after the glass slide 6, the sample support 1, and the
sample S are mounted on the support portion 12 of the mass
spectrometry device 10.
An example has been described in which the optical image of the
sample S and the sample support 1 is acquired in a state where the
sample support 1 is disposed on the sample S (the sixth step) is
performed after the distribution image is acquired (the fifth
step), but such acquisition of the optical image may be performed
at any time after the sample support 1 is disposed on the sample S
(the second step). For example, such an optical image may be
acquired before the glass slide 6, the sample support 1, and the
sample S are mounted on the support portion 12 of the mass
spectrometry device 10.
In the third step, as with the start point P1 and the end point P2,
the coordinates (X3,Y3) of the turn-around point P3 (refer to FIG.
6) may be grasped, and the coordinates of the turn-around point P3
of the irradiation range D2 may be recognized (stored), on the
basis of the position (x2,y1) of the sample stage 18 when the
markers 52 and 62 for a device corresponding to the coordinates
(X3,Y3) of the turn-around point P3 are read. In a case where there
is the information of the turn-around point P3, in addition to the
start point P1 and the end point P2, it is possible to more
accurately superimpose the optical image of the sample S on the
distribution image. For example, in a case where the optical image
is superimposed on the distribution image, and then, it is
necessary to match the direction such as an up-down direction and a
right-left direction, it is possible to suitably superimpose the
optical image on the distribution image, on the basis of the
information of three points not on a straight line (the start point
P1, the end point P2, and the turn-around point P3). In addition,
in a case where the irradiation range D2 is not in a rectangle
shape (for example, the irradiation range D2 is in a parallelogram
shape in which one side portion of four side portions intersects
with the Y axis direction), the information of the turn-around
point P3 may be read in a case where three or more reference points
(coordinates) are necessary for specifying the irradiation range
D2.
An example has been described in which the control unit 17 moves
the sample stage 18 such that the camera 16 scans the markers 52
and 62 for a device, but the scanning of the markers 52 and 62 for
a device by the camera 16 can be carried out by operating at least
one of the sample stage 18 and the camera 16. In a case where the
camera 16 is operated, the control unit 17 recognizes the
irradiation range D2, on the basis of the position of the camera 16
when the markers 52 and 62 for a device corresponding to the
irradiation range D2 determined by the measurer is read by the
camera 16. Accordingly, as with a case where the sample stage 18 is
operated, it is possible to accurately attain both of the
determination of the irradiation range D2 by the visual contact of
the measurer and the recognition of the irradiation range D2 by the
mechanical manipulation (the marker scanning) of the mass
spectrometry device 10, by the first marker 50 and the second
marker 60 provided in the frame 3.
REFERENCE SIGNS LIST
1: sample support, 2: substrate, 2a: first surface, 2b: second
surface, 2c: through hole, 3: frame (frame body), 4: conductive
layer, 10: mass spectrometry device, 13: irradiation unit, 16:
camera (scanning unit), 17: control unit, 31: first portion, 32:
second portion, 50: first marker, 51, 61: marker for visual
contact, 52, 62: marker for device, 60: second marker, D1: region
(existence range), D2: irradiation range, L: laser beam (energy
ray), R: effective region (ionization region), S: sample, S1:
component, S2: sample ion, w1, w4: width.
* * * * *